Note: Descriptions are shown in the official language in which they were submitted.
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MEDICATION FOR THE TR~ATMENT OR PRBVENTION OF HIV
VIRUS I~ lON THROUGH PASSIVE IMMUNIZATION AND PROCESSES
FOR ITS PREPARATION.
The present lnventlon relates to medlcatlon for the
treatment or preventlon of HIV vlrus lnfectlon, through
passlve lmmunlzatlon.
The use of speclflc antl-HIV gammaglobullns of human
orlgln obtalned from seroposltlve donors for the
seroprophylaxla and treatment of HIV (human
lmmunodeflclency vlrus) vlrus lnfectlon has been
contemplated prevlously. See Immunoglobulln preparatlon
for HIV-lnfected patlents - P.L. YAP and P.E. WILLIAMS -
Vos Sangulnls 55: 65-74 (1988).
In thls manner, PCT patent appllcatlon WO 89/01339
descrlbes lmmunoglobullns obtalned from healthy
seroposltlve donors and presentlng an lmportant p24
antlbody tlter.
However, these gammaglobullns contaln a wlde varlety
of dlfferent antl-HIV antlbodles, amongst whlch are
antlbodles to protelns of the vlral wall (ln the case of
HIVl: glycoprotelns gpl20 and gp41 or thelr precursor
gpl60), antlbodles to protelns of the vlral nucleus
(protelns p24, pl8 and pl5) or antlbodles to vlral
enzymes, partlcularly reverse transcrlptase (RT).
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Furthermore, seroposltlve donors could also hold
antibodles to the marker proteln of the T4 lymphocytes
(CD4 proteln) as a result of an auto-lmmunization
trlggered by the lnteractlon of vlral proteln gpl20 wlth
proteln CD4 whlch constltutes the natural receptor of
proteln gpl20 ln the T4 lymphocytes.
Now, antlbodles to the protelns of the vlral envelope
are suspected to "facllltate" lnstead of protect agalnst
vlral lnfectlon, apparently because they would form an
lmrnune complex wlth the vlrus and provoke lts phagocytosls
by macrophages, thus leadlng to an lnfectlon of the
macrophages whlch would consltute a reservolr for the HIV
vlrus.
Furthermore, antlbodles to the CD4 proteln are also
suspected to have a harmful effect durlng the HIV
lnfectlon by trlggerlng cytolysls of T4 lymphocytes,
thereby contrlbutlng to the development of the acqulred
lmmunodeflclency syndrome (AIDS), clinlcal manlfestatlon
of the HIV virus lnfectlon.
Also, lt has been observed that the outbreak of
clinlcal signs of AIDS on a prevlously asymptomatic
seroposltlve patlent ls accompanled by a collapse ln antl-
p24 and antl-RT antlbodles whereas the tlter of
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neutralizing antibodies in vitro is not modified. See:
Prospects for the control of AIDS by immunizing
seropositive individual: J. SALK, Nature 327: 473-76, June
11, 1987).
Other details on the HIV virus infection processes
are described in: Replication of the human
immunodeficiency virus - Strategies for inhibition. B.M.
PETERLIN and P.A. LUCIW, Biotechnology 6: 794-799, July
1988).
The present invention intends to provide medication
for the treatment or the prevention of the HIV virus
infection through the use of hyperimmune anti-HIV human
immunoglobilins.
The present invention therefore relates to medication
constituted by or comprising a preparation of hyperimmune
anti-HIV IgG and/or IgM gammaglobulins of human origin.
The preparation is devoid of antibodies to glycoproteins
of the HIV virus envelope and/or anti CD4 antibodies.
The medication of the present invention is
advantageously conditioned to be administered
parenterally, particularly through intramuscular or
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intravenous injection, in the same fashion as the usual
immunoglobulin preparations.
The present invention also relates to processes for
the preparation of the above-mentioned medication.
According to one embodiment of the processes of the
present invention, plasma from blood or placentas of HIV
virus seropositive donors or mothers is collected. From
this material, there is obtained, through known isolation
or separation processes, a total IgG and/or IgM
immunoglobulin preparation, and this preparation is
treated to eliminate anti-HIV envelope and/or anti CD4
antibodies.
In another embodiment of the processes of the present
invention, one or more antibodies to the HIV virus but not
to the envelope glycoproteins and/or CD4 are selectively
extracted from plasma, blood or placentas.
Particularly, one or more anti-core, anti-p24, anti-
p55, anti-pl8, anti reverse transcriptase or anti nef
antibodies may be extracted.
The extraction may, for example, be carried out by
immobilizing on a support (agarose gel, for example) one
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or more natural HIV antigens, or antigens bearing the
desired epitope, obtained through chemical synthesis or
expression in a suitable recombinant system.
One of the advantages conferred by the selective
extraction of the desired antibody from the collected
immunoglobulins is that higher concentration and
standardization of both the titer and the extracted
immunoglobulin composition from the collected
immunoglobulins are obtained. One may eventually mix the
selectively extracted immunoglobulins and obtain
standardized antibody titers.
When the anti-envelope and/or anti-CD4 antibodies are
selectively removed from the collected immunoglobulins,
it is preferable to repeat the removal steps several times
through extraction, in order to obtain an anti-envelope
and/or anti-CD4 antibody titer that is practically non-
existant.
If viral HIV particles are present in the material,
the process of the present invention also comprises a
further step through which the virus is removed or
inactivated.
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The seroposltlve donors or mothers may elther be
infected by the HIV vlrus or may have been vacclnated wlth
lnactlvated total HIV vlrus or HIV vlrus from a non-
lnfectlous straln.
For example, plasma, blood or placenta of HIVl
seroposltlve donors or mothers may be chosen but the
process of the present lnventlon may also be carrled out
uslng HIV2 or a mlxture of materlals related to HIVl and
HIV2.
Total IgG and/or IgM may be obtalned through
dlfferent known processes. For example, they may be
obtalned through COHN's alcohol separatlon process for
plasma or through TAYLOR's alcohol separatlon process for
placenta, or through chromatography. These separatlon
methods are well known to those skllled ln the art.
When the materlal has been obtalned from a
seropositlve donor lnfected by the wlld type vlrus, the
speclflc antl-HIV vlral lnactlvatlon treatment may, for
example, be obtalned through a beta-proplolactone
treatment, through exposure to heat or through an organlc
solvent ln the presence of a detergent. These
lnactlvatlon processes, whlch are well known to those
skllled ln the art, are thoroughly descrlbed ln processes
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for the preparation of immunoglobulins originating from
plasma or placenta. (See: Inactivation of the human
immunodeficiency viruses (HIV-l and HIV-2) during the
manufacturing of placental albumin and gammaglobulins. M.
GRANDGEORGE and F. PELLOGUIN, Transfusion 29: 629-634,
1989).
The removal of the anti-HIV envelope or anti CD4
antibodies may advantageously be carried out through bath
or column immunoadsorption by contacting the
immunoglobulin preparation with an insoluble support on
which envelope proteins gpl20 andtor gp41 and/or their
precursor gpl60 (so far as HIV1 is concerned) and/or
protein CD4 have been previously immobilized through any
method known to those skilled in the art. The proteins
may, for example, be immobilized on an agarose support
activated with cyanogen bromide. These proteins may be
obtained either from natural sources (HIV and lymphocytes)
or by genetic recombination.
According to another embodiment, plasma, blood or
placenta of HIV seropositive donors or mothers resulting
from vaccination with isolated HIV-virus proteins or
obtained by genetic recombination, with the exception of
the envelope proteins of the virus, may be utilized. In
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this process, the steps through which anti-envelope and/or
anti CD4 antibodies are eliminated are omitted.
The preparation obtained through one of the processes
referred to above is conditioned to be administered
intramuscularly or intravenously through classical
stabilization, additional purification, sterile filtration
and eventually lyophilization steps.
The medication of the present invention may be used
in the following applications:
- preventing HIV infection for persons susceptible of
contracting the disease through seroprophylaxis,
- adjuvant or complementary treatment of an anti-HIV
vaccination,
- treatment of the HIV infection and prevention of
its clinical manifestation, AIDS,
- treatment of AIDS.
Example 1.
PreParation of anti-HIV gammaglobulins concentrated in
anti-core antibodies and devoid of anti-enveloPe
antibodies.
The starting material is a preparation of human
placenta gammaglobulins, rich in HIV antibodies, obtained
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g
from a pool of HIV seroposltlve women placenta, purlfled
through an alcohol technlque completed by a viral
lnactlvatlon heatlng step. Thls gammaglobulln ls ad~usted
at a proteln concentratlon of 50g/lltre, ln a phosphate.
NaCl pH 7.4 buffer. The solutlon i8 tltrated ln ELIS.
(Envacor Abbott HIV test). The solutlon contalns a tlter
of:
l~/700 ln antl-core antlbodles
1/30 ln anti-envelope antibodies.
The titer is attrlbuted by the last posltlve dllutlon
ln the ELISA test.
- 80 ml of thls solutlon are flltrated on a column
contalnlng 3 mg of p25 antlgen immobilized on 1 ml of
agarose gel.
- The resulting filtrate contains essentially all the
desired gammaglobulins; ad~usted at 50 g/l in proteln, the
filtrate titers:
1/30 in anti-core antlbodles
1/30 ln antl-envelope antlbodleæ
The column ls then eluted wlth a glycocol acld
buffer.
Thls elutant contains 13 mg of protein, its tlter
belng brought back to a 50 g/l proteln concentratlon of:
1/120.000 ln antl-core antlbodles
negatlve in anti-envelope antibodieæ
r~
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This example illustrates the possibility of preparing
an anti-HIV globulin rich in anti-core antibody and
lacking anti-envelope antibodies.
Example 2
Preparation of an anti-HIV qammaglobulin lacking anti-
envelope antibodies.
The starting material used in this example is
identical to that used in example 1. The globulin is
adjusted at a protein concentration of 50 g/l in a
phosphate. NaCl pH 7.4 buffer.
The solution is titrated in ELISA (Envacor Abott HIV
test).
The solution has a titer
of 1/700 in anti-core antibodies
of 1/30 in anti-envelope antibodies.
- 60 ml of this solution are filtrated on a column
containing 4 mg of gpl60 antigen immobilized on 1 ml of
agarose gel.
- The filtrate collected contains essentially all the
proteins. Its titer, adjusted to 50 g/l in protein, is
the following:
1/500 in anti-core antibodies
1/8 in anti-envelope antibodies.
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A second run of the filtrate on the previously
regenerated column support allows the obtention of a
second filtrate which, when adjusted to 50 gtl, has the
following titer:
1/500 in anti-core antibodies
1/3 in anti-envelope antibodies.
After having carried out a third filtration on the
same support, using conditions similar to those set forth
above, the third filtrate obtained and adjusted at a
protein concentration of 50 g/l still contains most of the
proteins and shows the following titer:
1/500 in anti-core anbibodies
negative in anti-envelope antibodies.
This preparation illustrates the possibility of
preparing an anti-HIV gammaglobulin containing
particularly anti-core antibodies but lacking anti-
envelope antibodies.
